The gibbon ape leukemia virus is a family of type C retroviruses associated with several distinct hematopoietic malignancies. The Seato strain is associated with granulocytic leukemia. The principal determinant of enhancer activity has been shown to reside in a 22 bp element which interacts with cellular proteins. The gibbon T-cell lymphoma cell line MLA 144 strongly transactivates the GALV-Seato enhancer. Fractionation of MLA 144 extracts by conventional and affinity chromatography allows the separation of the GALV enhancer binding protein complex into 2 components. One of these components interacts strongly with DNA but does not in itself possess full specificity. The second component does not in itself bind DNA, but upon forming a complex with the first component confers greatly enhancer power to discriminate between different sequences. These proteins are distinct and separable from fos/jun (APl). However, a minor complex is present in MLA 144 which contains a fos-related antigen (FRA). The two components of the major complex can be independently activated in a cell-line specific manner. The first component has a molecular weight of 38-42,000 and is found most abundantly in T cells. The second component is smaller and ubiquitous. A variety of transfection and DNA-binding studies suggests that regulation of the interaction of components one and two may be an important control point for regulating T-cell activation.